The innovation of a peptide vaccine strategy may contribute to the development of efficacious and convenient cancer vaccines. Recently, we formulated an efficacious peptide vaccine without carriers using the natural phosphodiester bond CpG-DNA and a special liposome complex [Lipoplex(O)]. The peptide vaccine targeting a tumor antigen, transmembrane 4 superfamily member 5 protein (TM4SF5), was confirmed to have preventive and therapeutic effects in a mouse hepatocellular carcinoma (HCC) model. In this study, we demonstrated that the isotype-switched (IgM-IgD-) B cell population increased after immunization and that the functional memory response persisted for at least 70 days after the final immunization of mice. Delayed implantation of BNL-HCC cells significantly induced the peptide-specific IgG2a production in the immunized mice. Accordingly, tumor growth was inhibited and the survival rate increased. These results suggest that our peptide vaccine induces memory response, which is essential for cancer vaccine application.